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Method for preparing low molecular weight gamma-polyglutamic acid

A technology of polyglutamic acid and low molecular weight, which is applied in the field of preparation of low molecular weight γ-polyglutamic acid, can solve the problems of insufficient research and development investment, insufficient experience, restrictions on the popularization and application of γ-PGA, and reduce industrial costs and risks , Improve product recovery rate, enrich the effect of product types

Active Publication Date: 2015-07-29
SHANDONG FREDA BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Due to the complex preparation process and high production cost of low molecular weight γ-PGA, especially the separation, purification and refining process are obstacles to large-scale production, and many domestic R&D institutions and enterprises have insufficient experience in this area and insufficient R&D investment, resulting in product quality. Not high, the production capacity is very limited, which also limits the popularization and application of γ-PGA

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] (1) Fermentation broth preparation: Bacillus subtilis (Bacillus subtilis) FRD518CGMCC NO.6772 is fermented and cultivated to obtain a fermentation broth. The specific steps include:

[0027] a. Take 1 bacterial strain preserved in glycerol at -20°C in the refrigerator, inoculate a loop of seeds into a 500ml Erlenmeyer flask with 100ml of seed medium, and culture at 35-37.5°C with 200r / min shaker After 24 hours, the seed liquid was obtained, and the strain concentration reached OD 600 1.5~2.0;

[0028] b. Inoculate the prepared seed liquid into the fermentation medium, the inoculum amount is 2.5%, and culture with aeration and stirring at 35-37.5° C. for 48 hours to obtain a fermentation liquid, and the fermentation yield can reach 67 g / l.

[0029] Seed medium: glucose 10g / L, yeast extract 5g / L, ammonium sulfate 5g / L, sodium glutamate 10g / L, magnesium sulfate 1g / L, pH 7.2-7.5, sterilized at 115°C for 20min;

[0030] Fermentation medium: sucrose 40g / L, peptone 10g / L, ye...

Embodiment 2

[0041] (1) Fermentation broth preparation: Bacillus subtilis (Bacillus subtilis) FRD518CGMCC NO.6772 was fermented and cultivated to obtain a fermentation broth, and the specific steps were the same as in Example 1;

[0042] (2) Obtaining low-weight γ-polyglutamic acid:

[0043]Adjust the pH of the fermentation broth to 3.5, raise the temperature to 80°C, and keep it warm for 6 hours. When the temperature drops to about 60°C, add 1.5% activated carbon to decolorize for 75 minutes. Colorless material liquid; this process degrades γ-polyglutamic acid above 1000KDa in the fermentation broth into low molecular weight γ-polyglutamic acid with a molecular weight range of 10KDa-500KDa;

[0044] (3) Precipitation:

[0045] Adjust the pH of the above feed solution to 2.5 with hydrochloric acid, let it stand for 24 hours, precipitate out, filter the plate and frame to obtain a white powder, and wash it with purified water with a pH of 2.5 until the waste water is colorless and clear; ...

Embodiment 3

[0052] (1) Fermentation broth preparation: Bacillus subtilis (Bacillus subtilis) FRD518CGMCC NO.6772 was fermented and cultivated to obtain a fermentation broth, and the specific steps were the same as in Example 1;

[0053] (2) Obtaining low-weight γ-polyglutamic acid:

[0054] Adjust the pH of the fermentation broth to 4.0, raise the temperature to 85°C, and keep it warm for 5 hours. When the temperature drops to 60°C, add 2% activated carbon for decolorization for 60 minutes. Pigment liquid; this process degrades γ-polyglutamic acid above 1000KDa in the fermentation broth into low molecular weight γ-polyglutamic acid in the molecular weight range of 10KDa-500KDa;

[0055] (3) Precipitation:

[0056] Adjust the pH of the above feed solution to 3.0 with hydrochloric acid, let it stand for 24 hours, precipitate out, filter the plate and frame to obtain a white powder, and wash it with purified water with a pH of 3.0 until the waste water is colorless and clear;

[0057] (4) ...

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Abstract

The invention discloses a method for preparing low molecular weight gamma-polyglutamic acid. The method comprises the following steps: adjusting the pH value of a fermentation liquid to 3.5-4.0, degrading while keeping the temperature at 75-100 DEG C for 4-8 h to obtain the low molecular weight gamma-polyglutamic acid with the range of molecular weight being 10 KDa-500 KDa; adding activated carbon for decoloration and a plate frame for filtration after the temperature reduces to 0-60 DEG C, removing thallus, activated carbon and impurity protein to obtain clarified colorless feed liquid; adjusting the pH value of the feed liquid to 1.5-3.0 through hydrochloric acid, leaving the feed liquid to stand for 12-24 h, and separating out the precipitation; adding purified water to the white powder, and adjusting the pH value to 4.5-6.5, stirring the mixture for dissolution to obtain a solution with the concentration of 40% or above; then performing band drying and smashing to obtain the low molecular weight gamma-polyglutamic acid finished product. The yield of low molecular weight gamma-PGA prepared through the method reaches 85% or above and the purity reaches 95% or above, and the product quality is improved.

Description

technical field [0001] The invention relates to a method for preparing low molecular weight gamma-polyglutamic acid, which belongs to the technical field of industrial microorganisms. Background technique [0002] γ-polyglutamic acid (γ-PGA) is a high molecular amino acid polymer synthesized by microorganisms. It is non-polluting to the environment and is a green biological product. It has many unique physical, chemical and biological properties such as excellent biodegradability, film formation, fibrosis, plasticity, cohesiveness, and moisture retention. Today, with emphasis on environmental protection and sustainable development, this biosynthesized degradable functional material is favored by people and is gradually being used in pharmaceutical manufacturing, food processing, antifreeze and fresh preservation of vegetables, fruits, and seafood. It can also be developed and applied in many fields such as cosmetics industry, tobacco, leather manufacturing and plant seed pr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P13/02C12R1/125
Inventor 马双双苏移山张林军朱云峰李珍爱宗工理刘英梅赵文刚
Owner SHANDONG FREDA BIOTECH
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